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A rapid approach for isolating single fungal spores from rice blast diseased leaves

Liwang Fei: China Agricultural University

<div>Single spore isolation is a fundamental approach in plant pathology and microbiology to isolate and identify plant fungal pathogen from diseased field samples. However, routine single spore isolation procedure is time-consuming and has a high risk of contamination by other microorganisms. In this study, we developed a rapid approach for isolating single spore of the fungal pathogen, Magnaporthe oryzae, from rice blast diseased leaves in the field with low potential of contamination. Firstly, fresh diseased leaf with single lesion was directly pasted onto solid water agar plate in the field and was then maintained at four degree. Next, the single fungal spore attached to solid water agar was picked out with needle under stereomicroscope within up to one week, and then the agar plug with a single fungal spore was transferred onto solid oatmeal tomato agar plate for growth and preservation. According to our experience, a skilled technician could isolate over one hundred single spores in one working day with nearly no contamination. This new developed approach is also suitable for isolating single spore from diseased field samples caused by other fungi that produce aerial conidia.</div>